His Tag Antibody (359) - Azide and BSA Free

Simple Western: His Tag Antibody (359) [NBP2-43536] - Human purified protein. Simple Western image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: His Tag Antibody (359) [NBP2-43536] - Mock and transfected 293T cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: 6X His tag stained by 6X His tag antibody [359] diluted at 1:1000.Blue: Fluoroshield with DAPI.

Western Blot: His Tag Antibody (359) [NBP2-43536] - 6x His antibody detects 6x His protein by Western blot analysis. A. Non-induced cell lysates from E. coli expressing HIS-tagged proteins. B. IPTG-induced cell lysates from E. coli expressing HIS-tagged proteins. 15 % SDS-PAGE. 6x His antibody dilution: 1:5000.

Western Blot: His Tag Antibody (359) [NBP2-43536] - Analysis of cell lysate/extract of His-fused gene transfected HeLa cells A. 1/20 lysate/extract of transfected HeLa cells B. Control with 2 ug of preimmune mouse IgG C. Immunoprecipitation of His-fused protein by 2 ug of 6X His tag antibody [359] 12% SDS-PAGE The immunoprecipitated His-fused protein was detected by 6X His tag antibody [359] diluted at 1:1000.

Western Blot: His Tag Antibody (359) [NBP2-43536] - A) 2 ug GST-recombinant protein B) 2 ug His-recombinant protein C) 1 ug His-recombinant protein D) 0.5 ug His-recombinant protein E) 0.1 ug His-recombinant protein. 12% SDS-PAGE 6x His antibody at 1:5000.

ELISA: His Tag Antibody (359) [NBP2-43536] - 6X His tag antibody [359] detects His-tagged recombinant proteins by indirect ELISA analysis. Various dilutions of His-tagged recombinant proteins coated onto the 96-well plate were detected by using 0.5 ug/mL 6X His tag antibody [359] . Bound Abs were further detected by using HRP-conjugated goat anti-mouse IgG with HRP substrates.

Western Blot: His Tag Antibody (359) [NBP2-43536] - 6x His antibody detects 6x His protein by Western blot analysis.Lane A and B were IPTG-induced cell lysate from E. coli expressing different GST-tagged proteins. Lane C to G were IPTG induced cell lysate from E. coli expressing different His-tagged protein. 12 % SDS-PAGE6x His antibody (NBP2-43536) dilution: 1:5000

Western Blot: His Tag Antibody (359) [NBP2-43536] - Non-transfected (-) and transfected (+) 293T whole cell extracts (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with His Tag antibody [GT359] (NBP2-43536) diluted at 1:5000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

• Reactivity: ET
• Applications: WB, Simple Western, ELISA, ICC/IF, IP
• Clone: 359
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Azide and BSA Free

His Tag Antibody (359) - Azide and BSA Free Summary

• Description: Novus Biologicals Mouse His Tag Antibody (359) - Azide and BSA Free (NBP2-43536) is a monoclonal antibody validated for use in WB, ELISA, ICC/IF, Simple Western and IP. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: The immunogen used to generate this antibody corresponds to His Tag
• Isotype: IgG3
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000 - 1:10000
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:2000
  • Immunoprecipitation 1:100 - 1:500
  • Simple Western Validated by a verified customer review
  • Western Blot 1:1000 - 1:10000

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS, 20% Glycerol
• Preservative: No Preservative
• Purity: Protein G purified

Alternate Names for His Tag Antibody (359) - Azide and BSA Free

• 6 His epitope tag
• 6-His Tag
• 6X His Tag
• 6X His
• 6x-His Tag
• 6X-His
• H
• Hexa His tag
• HHHHHH epitope tag
• HHHHHH tag
• His Tag
• HIS
• His6-Tag
• polyhistidine tag
• polyHistidine
• Poly-histidine

Background

A his-tag (also known as histidine tag or polyhistidine tag) is a common epitope tag that typically consists of at least 6 histidine residues fused to either the carboxyl (C-) or amino (N-) terminus of a targeted recombinant protein to facilitate its purification and detection (1). The most common his-tag is the hexahistidine (His6/6-His) tag which has a theoretical molecular weight of 0.8kda (1). The histidine residues readily interact with transition metal ions such as Co2+, Ni2+, Cu2+, and Zn2+, making immobilized metal-affinity chromatography (IMAC) the preferred technique for his-tag purification (1, 2). Metal ions are immobilized and bound to by His-tags in the IMAC column via the histidine imidazole ring. The tagged protein can be eluted off the column by washing with buffers containing a low concentration of imidazole (1, 2). Due to its relatively small size, low immunogenicity, versatility under denaturing conditions, and minimal interference with the structure and function of proteins, the his-tag is one of the most widely used tags for protein purification (1-3).

References

1. Malhotra, A. (2009). Tagging for protein expression. Methods in Enzymology, Guide to Protein Purification, 2nd Edition, 463, 239-258. https://doi.org/10.1016/s0076-6879(09)63016-0

2. Terpe, K. (2003). Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523-533. https://doi.org/10.1007/s00253-002-1158-6

3. Booth, W. T., Schlachter, C. R., Pote, S., Ussin, N., Mank, N. J., Klapper, V., ... Chruszcz, M. (2018). Impact of an N-terminal polyhistidine tag on protein thermal stability. ACS Omega, 3(1), 760-768. https://doi.org/10.1021/acsomega.7b01598

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Review for His Tag Antibody (NBP2-43536) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer Simple Western Human 12/04/2018

Summary

• Application: Simple Western
• Sample Tested: Purified protein
• Species: Human
• Lot: 41290